环磷酰胺对以KLH为特异性抗原BALB/c小鼠TDAR实验定量分析

目的:利用环磷酰胺诱导BALB/c小鼠免疫功能异常动物模型,考察以血蓝蛋白(KLH)作为特异性抗原,T细胞依赖性抗体反应(TDAR)实验定量分析方法能否预测药物的免疫抑制作用。方法:平行对照组和环磷酰胺组给予注射环磷酰胺进行造模,阴性对照组不予处理;阴性对照组和环磷酰胺组经腹腔注射KLH进行免疫;应用流式技术分析外周血淋巴细胞亚群变化;采用间接ELISA法检测血清KLH IgG抗体含量;HE染色观察脾脏组织学改变。结果:进行免疫抑制的实验动物外周血CD3 +CD4 +、CD3 +CD8 +T淋巴细胞以及B淋巴细胞数量明显减少,与阴性对照相比差异具有统计学意义( P <0.05),同时脾脏内淋巴细胞数量减少;TDAR定量检测结果显示环磷酰胺能够降低外周血清KLH IgG抗体产生量,较阴性对照组相比差异具有统计学意义( P <0.05),而TDAR实验定性分析差异无统计学意义( P >0.05)。结论: TDAR实验定量分析能够很好地反映药物对机体的免疫抑制作用。

Quantitative analysis of TDAR in BALB/c mice with KLH as specific antigen by cyclophosphamide

Objective: To investigate whether quantitative analysis of TDAR test with KLH as specific antigen can predict the immunosuppression of BALB/c mice induced by cyclophosphamide. Methods: Parallel control group and cyclophosphamide group were injected with cyclophosphamide for modeling,while negative control group was not treated.Negative control group and cyclophosphamide group were immunized by intraperitoneal injection of KLH.Flow cytometry was used to analyze the changes of peripheral blood lymphocyte subsets.Indirect ELISA was used to detect serum KLH IgG antibody level.HE staining was used to observe the histological changes of spleen tissue. Results: The numbers of CD3 +CD4 +,CD3 +CD8 +T lymphocytes and B lymphocytes in peripheral blood of immunosuppressed experimental animals were significantly decreased compared with negative control group( P <0.05),and the numbers of lymphocytes in spleen were also decreased.Quantitative results of TDAR showed that cyclophosphamide could reduce the production of KLH IgG antibody in peripheral serum(compared with negative control group, P <0.05),but there was no statistical difference in qualitative analysis of TDAR test( P >0.05). Conclusion: Quantitative analysis of TDAR test can well reflect the immunosuppressive effect of drugs on the host.